Substituted 6-aminopenicillanic acids and salts



Patented Mar. 12, 1968 3,373,156 SUBSTITUTED G-AMINOPENICELANIC ACIDS AND SALTS Peter Bamberg, Berti] Ake Ekstriim, and Berndt Olof Harald Sjiiberg, Sodertalje, Sweden, assignors t Aktiebolaget Astra, Sodertalje, Sweden, a company of Sweden No Drawing. Filed Aug. 29, 1966, Ser. No. 575,533 Claims priority, application Sweden, Aug. 30, 1965,

13 Claims. (Cl. 260-2391) The present invention relates to antibiotic compounds and their preparation. In particular it concerns penicillins containing -a substituted N 2 sulfonylethoxycarbonylamino radical in the side chain.

The compounds prepared according to the present invention are of value as antibiotics, as nutritional supplement in animal feeds, as agents for treatment of mastitis in cattle, and as therapeutic agents in the treatment of infectious diseases in humans, animals and poultry.

It is an object of the present invention to obtain penici-llins containing a substituted 'N-2-sulfonylethoxycarbonylamino group in the side chain, and to provide a method for their production.

The present invention provides compounds of the general formula:

(JO-N CH-OO OH NHCOOR2SO2R: (I)

in which R is an alkylene group of not more than methylene radicals. One or more of the hydrogens on the R methylene radicals may be substituted by an aliphatic radical, a cycloaliphatic radical, an araliphatic radical, an aromatic radical, or a heterocyclic radical of not more than carbon atoms. All of the foregoing groups can be unsubstituted or substituted by one or more halogen atoms, lower alkyl groups, lower alkoxy groups, aryloxy groups, lower acyloxy groups, carbethoxy groups, lower acylamido groups, lower alkylmercapto groups, nitro groups, amino groups, substituted amino groups, or lower alkyl-or arylsubstituted sulfoxy groups;

R is an unsubstituted ethylene radical;

R is an aryl radical containing not more than 10 carbon atoms, which may be unsubstituted or substituted by one or more halogen atoms, lower alkyl radicals, lower alkoxy radicals, aryloxy radicals or nitro radicals;

R and R3 are illustrated by the 'following compounds:

N (2 p tolylsulphonylethoxycarbonyl)aminornethylpenicillin, 6 [D a (2 -p tolylsulphonylethoxycarbonylamino)phenylacetamido] penicillanic acid, on ('2 p tolylsulphonylethoxycarbonylamino) 2,4- dichlorobenzylpenicillin, a (2 p t-olylsulphonylethoxycarbonylamino) m fluorobenzyl penicillin, a (2 ptolylsulphonylethoxycarbonylamino) p methylbenzylpenicillin, a (2 p tolylsulphonylethoxyoarbonylamino) p dimethylaminobenzylpenicillin, a (2 ptolylsulphonylethoxycarbonylamino) o methoxy-benzylpenicillin, cc (2 p tolylsulphonylethoxycarbonylamino) p phenoxybenzylpenicillin, a (2 p tolylsulphonylethoxycarbonylamino) ,9 acetoxyethylpenicillin, oz (2 p tolylsulphonylethoxicarbonylamino)'ycarbethoxypropylpenicillin, oz (2 p tolylsulphonyleth-oxycarbonylamino) 'y methylsulpoxypropylpenicillin,

f3 (2 phenylsulphonylethoxycarbonylamino) oz 2- tienylethylpenicillin, a (2 phenylsulphonylethoxycarbonylamino) cyclohexylmethylpenicillin,

'y (2 p chlorophenylsulphonylethoxycarbonylamino) a phenylpropylpenicillin, cc (2 p chlorophenylsulphonylethoxycarbonylamin-o) 2 furylmethylpenicillin,

a 2 p ethoxyphenylsulphonylethoxycarbonylamino) 'y methylbutylpenicillin,

cc (2 p nitr-ophenylsulphonylethoxycarbonylamino)- ,B-phenylethylpenicillin.

The invention also comprises non-toxic salts of Formula I compounds. The salts include non-toxic metallic salts, such as sodium in case of a-(2-p-tolylsulphonylethoxycarbonylamino) m fluorobenzylpenici-llin, potassium, as in case of 6-[D-a-2-p-tolylsulphonylethoxycarbonylamino)-phenylacetamido]penicillanic acid, calcium, as in case of cz-(2-p-tolylsulphonylethoxycarbonylamino)- 2,4-dichlor0benzylpenicillin, aluminum, as in case of 0c- (2-p-tolylsulphonylethoxycarbonylamino) o rnethoxybenzylpenicillin, and non-toxic ammonium and substituted ammonium salts, such as the trialkylamines, including triethylamine, as in case of a-(2-p-tolylsulphonylethoxycarbonylamino)-ethylpenicillin, procaine, as in case of 0c (2 p ehlorophenylsulphonylethoxycarbonylamino)-B-methylpropylpenicillin, dibenzylamine, as in case of a (2 p-tolylsulphonylethoxycarbonylamino)18- acetoxypropylpenicillin, N-benzylphenethylamine, as in case of a-(2-p-tolylsulphonylethoxycarbonylamino)propylpenicillin, l-ephenamine, as in case of a-(2-p-chlorophenylsulphonylethoxycarbonylamino cyclopentylmethylpenicillin, N,N'-dibenzylethylenediamine, as in case of a- (2 p nitrophenylsulphonylethoxycarbonylamino) mchlorobenzylpenicillin, dehydroabiethylamine, as in case of a (2 phenylsulphonylethoxycarbonylamino)-benzylpenicillin, N,N-bisdehydroabiethylenediamine, as in case of 3-(2-p-tolylsulphonylethoxycarbonylamino)-a-phenylethylpenicillin and other amines, which have been used to form salts with benzylpenicillin or phenoxymethylpenicillin.

The present invention further provides a method for preparing Formula I compounds, which comprises reacting a compound of the general formula:

Rr-C OX I ia-o o 0-Rls 02-11, (H) or R SO -R -OCOCl with a compound of the general formula:

CH3 Y-NH-OH-O \CZOHQ o0-N'- oH-ooom (III) and CH3 \CZCHK CON CHCOOR4 NH: (I V) respectively, where R R and R have the same meaning as above. R is a hydrogen, a cation, or a dior trialkylsilyl group, such as a dimethylsilyl or a trimethylsilyl group, -COX and YNH are groups of atoms capable of reacting with each other with the formation of a carbon-nitrogen bond so that eventually upon hydrolysis, a --CONH-- group is obtained. Y is preferably a hydrogen atom or a dior trialky-lsilyl group, such as a dimethylsilyl or on trimethylsilyl group. a

The compound of general Formula 11 is an acid chlo- 7 ride or its functional equivalent such as an acid bromide,

an activated ester, an anhydride, a mixed anhydride, especially one formed with an alkoxy formic acid, or a derivative obtained by reaction between a carboxylic acid and a carbodijmide or other compounds functioning in the same way,'such as N,N'-carbonyldiimidazole or N- ethyI-5-phenylisoxazolium-3'-sulphonate. Formula II compounds are illustrated by (2-p-tolylsulphonylethoxy-carbonylamino)-acetyl chloride, ethoxyformic anhydride of oz (2 p tolysulphonylethoxycarbonylamino) phenylacetic acid, 2-p-tolylsulphonylethoxycarbonyl chloride, 2-

p-chlorophenylsulphonylethoxycarbonyl chloride.

chloroform, ethylacetate, butylacetate, benzene or toluene; or the acylation reaction can be performed in dry organic solvents such as methylene chloride or chloroform. Alternatively, the compound of the general Formula III is a N-trialkylsiylyaminopenicillanic acid, trialkylsilylester or a reaction product obtained by treatment of 6-aminopenicillam'c acid with silylating reagents such as N-trimethylsilyl-diethylamine .or dimethyldichlorosilane, capable of transferring dior trialkylsilyl groups to the acid. In these cases the acylation reaction is performed,

preferably in the presence of organic tertiary bases, in .dry homogeneous solutions of organic solvents such as ether, ,tetrahydrofurane, dioxane, dimethylformamide, N, N-dimethylacetamide, methylene chloride, chloroform, carbon tetrachloride, ethylacetate, benzene or toluene.

If the Formula II compounds contain one or more asymmetric centers the resulting Formula I compounds may exist in different diastereoisomeric forms, which are all antibacterially active.

It is understood that the present invention comprises the diastereoisomers as well as mixtures of them. i

In this specification and the claims the term lower designates at most 6 carbon atoms.

The following examples illustrate the invention, but in no way limit it:

Example 1. Preparati0n of N-Z-p-r0lylslllphenylethoxycarbon laminomethylpenicillin 6-aminopenicillanic acid (2.2 g., 0.01 mole) and triethylarnine (3 g., 0.03 mole) in 100 ml. of dry methylene chloride were stirred for 2 hours in an ice-bath. N-Z-ptolylsulphenylethoxycarbonylglycyl chloride (3.2 g., 0.01 mole) was added to the obtained light suspension and stirring and cooling was continued for 2 hours. Water (100 ml.) was added and the mixture was acidified to pH 2. After separation of the phase the organic layer was treated once more with 100 ml. of water at pH 2 and was then washed twice with water. The organic solution, containing the free penicillin, was extracted with N potassium bicarbonate solution till the aqueous phase was neutral. Freeze-drying of the latter gave 2.6 g. of the potassium salt of N-2-p-tolylsulphonylethoxycarbonylaminomethylpenicilliu with a purity of 88% (hydroxyl- A amine assay with 6-aminopenicillanic acid as standard). The product Was'found to inhibit the growth of Staph. aureus, Oxford at a concentration of 0.25 mcq./ml.

The N-Z-p-tolylsulphenylethoxycarbonylglycy chloride was prepared according to A. T. Kader and C. I. M. Stirling (J. Chem. Soc. 1962, 258).

Example 2.-Preparati0n of a-(2-p=tolylsulphenylethoxycarbonylamino)-heptylpenicillin a-(2 p -tolylsulphonylethoxycarbonylamino)-caprylic acid (6.7 g., 0.017 mole) and triethylamine (1.76 g., 0.017 mole) in 80 ml. of dimethylformamide was treated at -10 C. dropwise with ethyl chloroformate (1.9 g., 0.017 mole), while stirring. 10 min. after that all the chloroformate had been added an ice-cooled solution of ,6-aminopenicillanic acid (5.6 g., 0.026. mole) and triethylamine (2.9 g., 0.029 mole) Was rapidly added. Stirring was continued for 90 min. without external cooling. Water was added and the mixture was washed with ether, acidified to pH 2 and extracted twice with ether. The combined ether extracts were washed with water and then extracted with N potassium bicarbonate solution till the aqueous phase was neutral. Freeze-drying of the latter gave 7.6 g., of the potassium salt of ot-(N-2-p-tolylsulphonylethoxycarbonylamino)heptylpenicillin with ,a purity of 92% (hydroxylamine assay with the potassium salt of benzylpenicillin as standard).

The product was found to inhibit the growth of Staph.

' aureus Oxford at a concentration of 0.25 mcq./ml.

The starting material was prepared by treatment of' a-aminocaprylic acid methyl ester in chloroform with 2-p-tolyl sulphenylethyl chloroformate in presence of triethylamine. The obtained product was hydrolyzed by treating with acetic acid containing concentrated hydrochloric acid to give a-(2-p-tolylsulphenylethoxycarbonylamino)caprylic acid. For analysis it was converted into its cyclohexylamine salt, M.P. 117-120 C. (Found: C, 58.71; H, 8.13; N, 5.57%. Calculated forC- I-I N O S: C, 59.48; H, 8.32; N, 5.78%.)

formamide. Ethylchloroformate (0.87 g., 0.008 mole) was added dropwise with stirring at 10 C. 10 min. after that all of .the chloroformate had been added an ice-cooled solution of fi-aminopenicillanic acid (2.6 g., 0.012 mole) and triethylamine (1.4 g., 0.014 mole) in 30 ml. of Water was rapidly added and stirring was continued for 90 min. without external cooling. Water was added and the mixture was washed with ether, acidified to pH 2 and extracted twice with ether. The combined ether. extracts were washed with water and extracted with N potassium bicarbonate till the aqueous phase was neutral. Freeze-drying of the latter. gave 3.1 g. .of the potassium salt of G-[D-oc-(Z-P- tolylsulphonylethoxycarbonylamino) phenylacetamido] penicillanic acidwith a purity of 75% (hydroxylamine assay with the potassium salt of benzylpenicillin as stand? ard) The product was found to inhibitthe growth of Staph.

' aureus Oxford at a .concentration of 0.25 meg/ml.

' traction with .ether at pH 2. For analysis it was converted into its cyclohexylamine salt, M.P. 162-163 C. (Found: C, 60.7 H, 6.65; N, 6.17; S, 6.18%. Calculated for C H N OGS: C, 60.48; H, 6.77; N, 5.88 S, 6.73%.)

In the manner, described in Examples 2 and 3, the

' following N-2 p tolylsulphonylethoxycarbonylaminopenicillins were prepared. In the table are given: yields and purities (hydroxylamine assay) of the products together with their activities against Staph. aureus Oxford. (M.I.C.=minimum inhibitory concentration in mcg./ml., determined in serial dilution tests on agar plates. [TSEC means 2-p-tolylsulphonylethoxycarbonyl.])

Penicillin Yield Purity M.I.C.

(Percent) (Percent) (meg/ml.)

a-TSE Ca.minopr0py1penici1ljn 22 89 a-TSE C-aminobutylpenicillin 48 73 0. 25 a-TSEC-aminopentylpem'cillin 60 64 0. 25 a-TSEC-aminomethylbutylpenicillin 46 80 0. 63 a-TsEC-aminofi-methylbutylpenicillin 34 56 0. 63 a-TSE C-amino-y-thiomethylpropylpenicillin 47 68 0. 13 6 -(L-oz-TSE C-amino-fJ-methylbutyramido) penicillinnic acid 45 68 6-[L-a-TSEC-amino-(L)-B-methylvaleramido] penicillauic acid 57 66 1. 25 fi-(L-cz-TSE Gamino-y-thio-rnethflbutyramido) panicillanic acid 22 66 fi-(D-az-TSE C-amino-y-thio-methylbutyramido) penicillanic acid 47 85 0. 63 6TSE C-aminobutylpenieilliu.-. 9 38 0. 63

The N-2 p-tolylsulphonylethoxycarbonylamino acids used as starting material for the penicillins, mentioned above, were prepared by treatment of the aminoacids in aqueous solutions at pH 9 with 2-p-toly1sulphonylethoxycarbonyl chloride as described in Example 3. Alternative 1y they were prepared via their esters as described in Example 2. Usually cyclohexylamine or dicyclohexylamiue salts of the acids were prepared for analysis. The following physical were recorded data:

The D-rx-(2 phenylsulphonylethoxycarbonylamino)- phenylacetic acid was obtained by treatment of 13-0:- aminophenylacetic acid in aqueous solution at pH 9 with 2-phenylsulphonylethoxycarbonyl chloride. The product was isolated by extraction with ether at pH 2, and was crystallized from ethylacetate-ligroin, M.P. 159-161 C. (Found: C, 56.80; H, 4.79; N, 3.64; S, 8.81. Calc. for C H NO S: C, 56.18; H, 4.72; N, 3.86; S, 8.82.)

Example 5.-Preparazi0n of 6-[u-(2-p-t0lylsulphonylethoxycarbonylamino)-fl-methyl-fi-(p-methoxyphenyl)butyramido] pencillanic acid or (2 p tolylsulphonylethoxycarbonylamino) flmethyl-,6-(p-methoxyphenyl)butyric acid (2.1 g. 0.0045 mole) and triethylamine (0.45 g., 0.0045 mole) were stirred in 25 ml. of dry dimethylformamide and treated dropwise at -l0 C. with isobutyl chloroformate (0.61 g., 0.0045 mole). 10 min. after that all of the chloroforrnate had been added a solution, in 10 ml. of dry dimethylformamide, of a trimethylsilyl-derivate of 6-aminopenicillanic acid (prepared as described in British Patent No. 959,853, Example 8, from 1.5 g. (0.0063 mole) of 6-aminopenicillanic acid) and N-trimethylsilylamine, was added and stirring was continued for 90 min. without external cooling. The reaction mixture was poured into water, washed with ether, acidified and extracted twice with ether. The combined ether extracts were washed with water and extracted witd N potassium bicarbonate till the aqueous phase was neutral. Freeze-drying of the latter N-z- -toi lsul honyl-e Elemental analysis (percent) carbonyl compound ot- M.P. C.)

C H N S a-Aminobutyric acid 1 128-130 Found 56. 94 7. 97 6. 52 Cale 57. 87 7. 95 6.14 Norvaline 1 C HQ-nN OaS 118-120 Found 56. 80 7. 78 6. 29 a1 56. 99 7. 74 6. 33 L-valine CHz1NOeS 108-113 52. 2 6. 10 4. 00 52. 46 6. 16 4. 08 N orleucine CzzHaaNgOaS 122-128 56. 88 7. 81 6. 51 57. 87 7. 95 6. 14 L-isoleucine CzzHaaNzOgS 150-152 57. 7 7. 84 6. 15 57. 87 7. 95 6. 14 Leueine l CzzHgaNgOaS 142-145 57. 00 7. 88 6. 27 g 57. 87 7. 95 6. 14 Alloisoleucine 1 CzzNaeNgOaS 128-130 56. 94 7. 97 6. 52 57. 87 7. 95 6. 14 III-methionine} CZ1H34NZO5SQ 129-132 51. 9 7. 6. 11 53. 14 7. 22 5. 90 Methionine 2 C27H44N305S7 141-144 57. 5 7. 75 5. 31 58. 2 7. 97 5. 03 D-methionine 1 CmHzrNgOeSg 124-126 52. 31 7. 13 5. 95 53. 14 7. 22 5. 90 E-Aminovaleric acid C1 Hz1N 05S 70- 79 52. 3 6. 27 3. 96 52. 46 6. 16 4. 08

1 Cyclohexylamine salt. Dicyclohexylarnine salt.

Example 4.Preparati0n of 6-[D-rx-(2-phenylsulph0nylethoxycarbonyl-amino)-phenylacetamia'o] penicillanic acid D-oz-(Z phenylsulphonylethoxycarbonylamino)phenylacetic acid (1.75 g., 0.0048 mole) was dissolved together with triethylamine (0.49 g., 0.0048 mole) in ml. of dry dimethylforma-mide. Ethyl chloroformate (0.52 g., 0.0048 mole) was added dropwise with stirring at 10 C. 10 'min. after that all of the chloroformate had been added a chilled solution of 6-aminopenicillanic acid (1.6 g., 0.0072 mole) and triethylamine (0.78 g., 0.0077 mole) was rapidly added and stirring was continued for 90 min. without external cooling. Water was added and the reaction mixture was washed with ether, acidified to pH 2 and extracted twice with ether. The combined ether layers were washed with water and extracted with potassium Jbic'arbonate solution till the aqueous phase was neutral.

Freeze-drying of it gave 1.9 g. of the potassium salt of 6-[D oz (2 phenylsulphonylethoxycarbonylamino) phenylacetamido] pencillanic acid with a purity of 92% (hydroxylamine assay).

The product was found to inhibit the growth of Staph.

aureus Oxford at a concentration of 0.13 meg/ml.

Example 6.Preparati0n of 6-[d(Z-p-tolylsulplzonylethoxycarbonylamino) -a-phenylpropionamido] penicillanic acid To a stirred and cooled solution of B-amino-ot-phenylpropionic acid (3.3 g., 0.02 mole) in 50% dic-xane at pH 9, 2-p-tolylsulphonylethoxycarbonyl chloride (5.8 g.,

0.0022 mole), dissolved in 10 ml. of benzene, was added dropwise while keeping the pH constant by addition of 2 N sodium hydroxide. After the consumption of alkali bined ether extracts were washed with water, dried and evaporated to give {3-(2-p-tolylsulphonylethoxycarbonylminimum inhibitory concentartion in mcg./ml., determined in serial dilution tests on agar plates). TSE means 2-p-tolylsulphonylethoxycarbonyl.

arnino)-u-phenyl propionic acid (6.3 g.) as a slightly 5 Penicillin Yield Purity 'M- coloured oil. This was dissolved together with triethyl- (percent) (percent) amine (1.7 g., 0.017 mole) in 100 ml. of dry dimethyl- -a -E-a9et formamide and treated dropwise with ethyl chloroforrnate 8 6 (1.85 g., 0.017 mole), while stirring at C. 10 min. f l 2 V 6 2 after that all of the chloroformate had been added an ice- 1O fi ig iggifilfifil fifiggga: 8 58 5 cooled solution of 6-aminopenicillanic acid (4.3 g., 0.02 gg a fk 26 75 (L mole) and triethylarnine (2.2 g., 0.022 mole) in 50 ml. penieillin 60 64 0.63

of water was added and stirring was continued for 90 82 0'13 mm. without external coohng. Water was added and the r il l n l figrsn m 77 O 25 reaction mixture was washed with ether acidified to pH 9 9t 171161110} -"ISEC-am111 1 2 and extracted twice wlth ether. The combined ether m 3 0.25- extractswere washed with water and extracted with a N potassium bicarbonate solution till the aqueous phase The corresponding N-2p-tolylsulphonylethoxycarbo- Was neutral. Freeze-drying of the latter gave 6.2 g. of nylamino acids were prepared by treatment of the amino the potassium salt of 6-[,8-(2-p-tolylsulphonylethoxycar- 20 acids in aqueous solutions at pH 9 with 2-p-tolyl su lbonylamino) cc phenylpropi amido]pcnicillanic acid phonylethoxycarbonyl chloride as describedin Example with a purity of 61% (hydroxylamine assay with the 3. Alternatively they were prepared via their esters as potassium salt of benzylpenicillin as standard). described in Example 2. In one case a cyclohexylamine The product was found to inhibit the growth of Staph. salt of the acid was prepared for analysis. The followaureus Oxford at a concentration of 0.25 meg/ml. 25 ing physical data were recorded.

N-2-p-tolylsulphony1- Elemental analysis (percent) ethoxycarbonyl M.P. 0.)

compound of C H N S Z-N-acetyl-L-lysine CmElnNzOrS 139440 Methionine sulphoxide C15H21NO1S: 137-138' a-an 1(ii.r1 o-m-nitrophenylacetic Cz-tHsrNaOaS 167 8.01 fl-arnino-a-(2-thienyl)proplonic C17H1 NOoS2 121-122 fi-gii liino-ww-thienyl)-pr0pionic CHHQNOISSH 151-152 i ininobut ne acid CHHwNOuS 103-110 1 Oyclohexylamine salt.

Example 7.-Preparati0n of 6-[ -(Z-p-tolylsulphonyb Example 8.Preparati0n of u-(Z-phefiylsulplionylethoxyethoaycarbonylamino)-bu!ymmid0]-pei1icillarzic acid carbonylamino)-'y-methylthiopropylpen icillin -(2-p-to1ylsu1phonylethoxycarbonylamino)butyric acid N 2 phenylsnlphonylethoxycarbonylmethionine (7 g., V (5.1 g., 0.016 mole) and triethylamine (1.6 g., 0.016 0.02 mole) was dissolved in dry dimethylfor mamide (150 mole) in 100 ml. of dry dimethylforrnamide were stirred ml.), containing triethylamine (2 g., 0.02 mole). Ethyl at 10 C. and treated dropwise with ethyl chloroforrnchloroformate (2.2 g., 0.02 mole) was added dropwise ate (1.7 g., 0.016 mole). 10 min. after that the addition with stirring at 10 C. 10 min. after that all of the chloof the latter was complete, an ice-cooled solution of 6- roformate had been added a chilled solution of d-aminoaminopenicillanic acid (5.1 g., 0.023 mole) and triethylpenicillanic acid (6.5 g., 0.03 mole) and triethylarnine amine (2.5 g., 0.025 mole) in 60 ml. of water was rap- (3.2 g., 0.032 mole) in water (35 ml.) was rapidly added idly added and stirring was continued for 90 min. withand stirring was continued for 20 min. without external out external cooling. Water was added and the reaction cooling. Water was added and the reaction mixture was mixture was washed with ether, acidified to pH 2 and washed with ether, acidified to pH 2 and extracted twice extracted twice with ether. The combined ether extracts with ether. The combined ether extracts were washed with were washed with water and extracted with N potassium water and extracted with N potassium bicarbonate solubicarbonate till the aqueous phase was neutral. Freezetion till the aqueous phase was neutral. Freeze-drying of drying of the latter gave 0.7 got the potassium salt of it gave 1.4 g. of penicillin with apurity of 77% (hydroxyl- 6 (2-p-tolylsulphonylethoxycarbonyla'mino)-penicilamine assay with benzylpenicillin as standard). lanie acid with a purity of 35% (hydroxylamine assay The N-2-phenylsulphonylethoxycarbonylmethionine with the potassium salt of benzylpenicillin as standard). was obtained by treatment of methionine in aqueous solu- This product was found to inhibit the growth of Staph. tion at pH 9 with 2-phenyl-sulphonylethoxycarbonyl ch10- anreus Oxford at a concentration of 0.25 meg/ml. ride. The product was isolated by extraction with ether at a 'y (2 p tolylsulphonylethoxycarbonylarnino)butyric pH 2. For analysis the cyclohexylamine salt was prepared, acid, M.P. 106109 C., was obtained by treatment of M.P. 12026 C. (Found: C, 52.33; H, 7.11; N, 5.88; S,

'y-arninobutyric acid at pH 9 in 50% dioxane with 2-p- 13.81. Calc. for C H N O S C, 52.15; H, 7. 00; N, 6.08; tolylsulphonylethoxycarbonyl chloride. It was isolated by S, 13.92%). enxgziton With ether after acidification of the reactlon Example grqprepamtion of w(z p chlomphmylsulphom In the manna. described in Examples 2 and 3 the ylethoxycarbonylamzno)-'y-methylthzopropylpemczllm lowing N-2-p-tolylsulphonylethoxycarbonylaminopenicil- N 2 p chlorophenylsulphonylethoxycarbonyl-methlins were prepared. In the table are given yields and ur- V ionine (5.9 g., 0.015 mole) and triethylamine (1.5 g., ities (hydroxylamine assay) of the productstogether with 0.015 mole) in dry dirnethylforrnamide ml.) were their activities against Staph. auerus Oxford (M.I.C.= 75

stirred at 10 C.- -15 (3., while ethyl chloroformate (1.6 g., 0.015 mole) dissolved in a few ml. of ether, was added dropwise. 10 min. after that all of the chloroformate had been added an ice-cold solution of 6-aminopenicillanic acid (4.2 g., 0.023 mole) and triethylamine (2.4 g., 0.024 mole) in water (65 ml.) was rapidly added and stirring was continued for 90 min. without external cooling. Water was added and the mixture was washed with ether, acidified to pH 1.5 and extracted with ether. The combined ether extracts were washed with water and extracted with N potassium bicarbonate till the aqueous phase was neutral. Freeze-drying of the latter gave 7.5 g. of the potassium salt u-(2-p-chlorophenylsulphonylethoxycarbonylamino -'y-methylthiopropylpenicillin with a purity of 77% (hydroxylamine assay with benzylpenicillin as standard).

The product was found to inhibit the growth of Staph. anreus Oxford at a concentration of 0.25 meg/ml.

The N 2 p chlorophenylsulphonylethoxycarbonylmethionine, M.P. 9597 C. (Found: C, 42.71; H, 4.56; CI, 8.96; N, 3.53; S, 16.02. Calc. for C H ClNO S C, 42.47; H, 4.58; Cl, 8.96; N, 3.54; S, 16.20%) was prepared by treatment of methionine in aqueous solution at pH 9 with 2-p-chlorophenylsulphonylethoxycarbonyl chloride. The product was isolated by extraction at pH 2 and recrystallized from ethyl acetate.

In the Way, described in Example 9 the a-(Z-p-chlorophenylsulphonylethoxycarbonylamino) -methylsulphonyl-propylpenicillin was prepared.

Example 10.-Prepnratin of 6[D-a-(Z-p-tolylsulphonylerhoxycarbonylamz'no)-phenylacetamido]penicillanic acid 6-(D-a-amino-phenylacetamido)penicillanic acid (85% pure, 41 g., 0.1 mole) suspended in a mixture of water (25 0 ml.) and methylisobutylketone (250 ml.) was treated at pH 8.5 with 2-p-tolylsulphonylethoxycarbonyl chloride (97% pure, 27.2 g., 0.1 mole), dissolved in dry ether (100 ml.), while stirring and cooling in an ice-bath. The pH of the reaction mixture was kept constant by addition of 2 N sodium hydroxide. After the consumption of base had ceased, the reaction mixture was washed with ether and butylacetate, acidified to pH 1.5 and extracted several times with butylacetate. The combined butylacetate extracts were washed with water, dried over anhydrous magnesium sulphate for 45 min. and filtered. Isopropylamine (5.9 g., 0.1 mole) was added and a crystalline precipitate of the isopropylamine salt of 6-[D-2-(2-p-tolylsulphonylethoxycarbonylamino)-phenylacetamido] -penicillanic acid was formed on cooling. After standing three days in the icebox, it was filtered, washed and dried. There was obtained 60.8 g. of the penicillin with a purity of 94% (hydroxylamine assay with benzylpenicillin as standard).

Example 11.Preparati0n 0] 6-[D-w(2-p-chlorophenylsulphonyloxycarbonylamino) phenylacetamido]penicillanic acid 6 (D a amino phenylacetamido)penicillanic acid (85% pure, 20.7 g., 0.05 mole) in a stirred and cooled mixture of water (150 ml.) and methylisobutylketone 150 ml.) was treated at pH 8.5 with 2-p-chlorophenylsulphonylethoxycarbonyl chloride (14.2 g., 0.05 mole), dissolved in dry ether. After the reaction was complete, the mixture was washed with ether, acidified to pH 1.5 and extracted with ether repeatedly. The combined ether extracts were washed with water and extracted with N potassium bicarbonate solution (150 ml). The pH of the aqueous phase was adjusted to 6.5 and the solution was freeze-dried to give 28.1 g. of the potassium salt of 6- [D oz 4 (2 -p-chlorophenylsulphonylethoxycarbonylamino)-phenylacetamido]penicillanic acid with a purity of 74% (hydroxylaniine assay with benzylpenicillin as standard). Paper chromatography showed that the product contained minor amounts of 6-(D-a-amino-phenylacetamido)penicillanic acid and of a further compound with antibiotic properties as lay-products.

. 10 Example 12.Preparation of 6-[D-a-(Z-p-nilrophenylsulphonylethoxy carbonyl amin0)-phenylacetamid0] penicillanz'c acid A stirred and cooled mixture of 6-(D-u-aminophenylacetamido)-penicillanic acid (84%, 9.7 g., 0.023 mole), methylisobutylketone (70 ml.) and water (70 ml.) was treated with 2 p nitrophenylsulphonylethoxycarbonyl chloride (6.9 g., 0.023 mole), while the pH of the reaction mixture was kept constant at 8.5 by addition of 2 N sodium hydroxide. After the consumption of base had ceased the reaction mixture was washed with ethyl acetate, acidified to pH 1.5 and extracted with ethylacetate. The combined ethylacetate extracts were washed with Water and extracted with N potassium bicarbonate till the aqueous phase was neutral. After washing with ether the aqueous phase was adjusted to pH 6.5 and pure-dried to give 5.8 g. of the potassium salt of 6-[-a-(2-p-nitrophenylsulphonylethoxycarbonylamino) phenylacetamido] penicillanic acid and with a purity of 64% (hydroxylamine assay with benzylpenicillin as standard).

We claim:

1. Penicillins having the formula:

and non-toxic salts thereof; wherein R is selected from the group consisting of unsubstituted and substituted lower alkyl radicals, with the substituted lower alkyl radicals having substituent radicals selected from the group consisting of lower alkyl, lower alkanoyloxy, monocarbocyclic aryl containing from 6 to 10 carbon atoms, carbethoxy, lower alkanoyloxyamino and lower monocarbocyclic aryloxyamino, lower alkyl sulfoxy, lower alkyl sulfonyl, lower alkyl thio and sulfur and oxygensubstituted monohetercyclic radicals containing a maxi mum of 10 carbon atoms, and monocarbocyclic aryloxy radicals containing from 6 to 10 carbon atoms; R is ethylene; R is selected from the group consisting of unsubstituted and substituted monocarbocyclic aryl radicals containing from 6 to 10 carbon atoms, with the substituted radicals having substituent radicals selected from the group consisting of halide, lower alkyl, lower alkoxy, and nitro radicals.

2. Penicillins having the formula:

and non-toxic salts thereof; wherein R is selected from the group consisting of unsubstituted and substituted benzyl radicals, with the substituted lower alkyl radicals having substituent radicals selected from the group consisting of halide, lower alkyl, lower alkyla-mino, nitro, lower alkoxy, monocarbocyclic aryloxy containing from 6 to 10 carbon atoms; monocarbocyclic aryl radicals containing from 6 to 10 carbon atoms and monocarbocyclic aryloxy radicals containing from 6 to 10 carbon atoms; R is ethylene; R is selected from the group consisting of unsubstituted and substituted monocarbocyclic aryl radicals containing from 6 to 10 carbon atoms, with the substituted radicals having substituent radicals selected from the group consisting of halide, lower alkyl, lower alkoxy, and nitro radicals.

3. Compounds as set forth in claim 2, in which R is an unsubstituted benzyl radical.

4. Compounds as set forth in claim 1, wherein R is an a-thienylethyl radical with the thienyl group in a position selected from the group consisting of positions two and three.

1 1 1 Z 5. Compounds as set forth in claim 1, wherein R is 11. A compound as set forth in claim 1; which is 6-[,8- p-tolyl. (2 p tolylsulfonylethoxycarbonylamino)-a-phcnylpro- 6. Compounds as set forth in claim 1, wherein R is pionamido] penicillanic acid. phenyl. 12. Compounds as set forth in claim 2, wherein R is 7. A compound as set forth in claim 1, which is N 2-p- 5 p-tolyl. tolylsulfonylethoxycarbonylaminomethyl penicillin. 13. Compounds as set forth in claim 2, wherein R is 8. A compound as set forth in claim 1, which is Oc-(Z-pphenyl. tolylsulfon-ylethoxycarbonylamino)heptyl penicillin. References Cited 9. A compound as set forth in claim 1, which is 6-[D- UNITED STATES PATENTS a (2 p-tolylsulfonylethoxycarbonylamino) henylacetamide] penicillanic acid.

10. A compound as set forth in claim 1, which is 6-[a- (2 p tolylsulfonylethoxycarbonylamino) fi-methylfi- NICHOLAS Pnmary Exammer' (p-methoxyphenyl)butyramido] penicillanic acid.

10 3,245,983 I 4/1966 Doyle et al. 260-239 UNITED STATES PATENT OFFICE CERTIFICATE OF CORRECTION Patent No 3,373 ,156 March 12 1968 Peter Bamberg et a1 It is hereby certified that error appears in the above numbered patent requiring correction and that the said Letters Patent should read as corrected below.

Column 2, lines 4 and 5, "methylsulpoxypropylpenici1lin" should read methylsulphoxypropylpenicillin Column 3, line 37, "N-trialk'ylsiylyaminopenicillanic" should read N-trialkylsilylaminopenicillanic lines 59 and 60, "N-Z-p-tolylsulphenyletho'xycarbonylaminomethylpenicillin" should read N-2 p-tolylsulphonylethoxycarbonylaminomethylpenicill-in line: 63 and 64, N-'2-ptolylsulphenylethoxycarbonylglycyl" should read N-2-p tol'ylsulphonylethoxycarbonylglycyl Column 4, lines 3 and 29', "mcq", each occurrence, should read mcg line 4, "N'Z*p-'tolylsulphenylethoxycarbonylglycy" should read H N 2-p-tolylsulphonylethoxycarbonylglycyl line 7 oc- (2ptolYlsulphenyleth'Oxy-"" should read or (2*p-to1ylsu1phony1- ethoxy line 32", "2-p-to1y1sulphenylethyl" should read T-= 2 -p to1y1su1phonylethy1" P lines 35" and 36; c- (2-ptolylsulphenylethoxycarbonylamino) should read a u- (2-p- Ic olylsulphonyle'thoxycarbonylamino) Column 5, line 44,

,, I o to Y C N N O S should read C H N O S Column 6, line 28, "witd" should read with line 73, "0.0022" should read 0.022 Column 7, line 60, after "(Z-p-tolylsulphonylethoxycarbonylamino)-" insert butyramido] Column 10,

line 36, "alkan'oyloxyamino" should read alkanoylamino line 37, "aryloxyamino" should read H aroylamino line 57 "lower alkyl" should read benzyl Signed and sealed this 12th day of August 1969.

(SEAL) Attest:

EDWARD M.FLETCHER,JR. WILLIAM E. SCHUYLER, JR. Attesting Officer Commissioner of Patents 

1. PENICILLINS HAVING THE FORMULA: 